【摘要】 目的：观察糖尿病大鼠视网膜病变,探讨牛磺酸对糖尿病大鼠VEGF，TSP1的影响。方法：选择健康成年SD大鼠30只,随机分成正常对照组，糖尿病组，牛磺酸组。一次性腹腔内注射链尿佐菌素(STZ),建立糖尿病大鼠模型,在普通饲料中加12g/L牛磺酸进行营养干预,采用免疫组化SABC法，对视网膜上VEGF，TSP1进行研究。对各组视网膜因子表达情况进行图像分析。结果：与糖尿病组比较,牛磺酸组视网膜VEGF蛋白质的阳性表达降低；TSP1蛋白质阳性表达升高,差别显著。结论：牛磺酸可明显改善糖尿病视网膜的VEGF，TSP1的表达。
【关键词】 牛磺酸；糖尿病视网膜；VEGF；TSP1
Effect of taurine on VEGF, TSP1 of rats with earlystage diabetic retinopathy
DaiYuan Shi, Mu Li, ZhiWei Chen
1Department of Ophthalmology, Central Hospitall, Gaizhou 115200, Liaoning Province, China;
2Department of Ophthalmology, Eye Hospital, Liaoyang 111000, Liaoning Province, China;
3Qiqihar Medical College, Qiqihar 161006, Heilongjiang Province, China
AbstractAIM: To observe retinopathy of diabetic rats and to investigate the effect of taurine on VEGF, TSP1 of rats with earlystage diabetic retinopathy.METHODS: Thirty SD rats were chosen and randomly divided into normal group, diabetic group and taurinetreated diabetic group. Diabetic rats were induced by injected streptozocin into abdominal cavity once and were treated with taurine (12g/L) that were mixed with food. Three months after the models being created, retinal tissues of rats were observed with optical microscope, SABC to detect WEGF and TSP1.RESULTS: Compared with diabetic group, the expression of WEGF was obviously decreased, while TSP1 expression was significantly increased in taurinetreated diabetic group.CONCLUSION: Taurine can effectively protect retinal capillaries and improve the expression of WEGF and TSP1.
KEYWORDS: taurine; diabetic retina; WEGF; TSP1
0引言
糖尿病视网膜病变(diabetic retinopathy,DR)是糖尿病的主要并发症之一,几乎所有糖尿病病程达15a以上的患者均出现一定程度的视网膜病变,牛磺酸是视网膜中含量最丰富的游离氨基酸,是视网膜发育必需的营养因子,研究人员发现糖尿病视网膜中牛磺酸含量减少,补充牛磺酸可以防治糖尿病视网膜病变。为探讨牛磺酸对糖尿病视网膜病变的防治作用,我们观察了其对糖尿病视网膜VEGF，TSP1的影响。
1材料和方法
1.1材料
实验用封闭群SD大鼠30只,雌雄不限，体质量220～250g,自然昼夜光线照明。动物随机分为3组：正常对照组、糖尿病组、牛磺酸组，每组10只。大鼠正常组及糖尿病组饲以标准饲料,牛磺酸干预组的饲料在标准饲料中加入12g/L牛磺酸,牛磺酸购自北京世纪维他保健品公司;兔抗小鼠VEGF单克隆抗体中山生物技术有限公司,山羊抗小鼠TSP1单克隆抗体美国NeoMarkers公司,阳性对照片：为人乳腺癌的VEGF和TSP1免疫组化染色片,中山生物技术有限公司,SP9002免疫组织化学试剂盒中山生物技术有限公司,DAB显色试剂盒武汉博士德生物技术有限公司。
1.2方法
实验前大鼠禁食12h,自由饮水用链脲佐菌素(Streptozotocin,STZ,Sigma公司)诱导糖尿病,临用前用0.1mmol/L的柠檬酸缓冲液(pH4.5)配制成10g/L的STZ溶液,按60mg/kg一次性左下腹腔注射,注射后72h检测定性尿糖和血糖,用尿糖试纸测尿糖达+++以上,尾静脉采血测血糖浓度>6.7mmol/L即为模型建立成功[1]。实验期间测血糖，1次/mo,各组动物3mo后，4℃预冷的40g/L聚甲醛灌注固定大鼠,取大鼠左眼球于40g/L多聚甲醛后固定4～6h,300g/L蔗糖4℃过夜,冰冻切片将视网膜中部(即贯穿视神经)的切片取3张,采用免疫组化SABC法测定VEGF，TSP1的表达。
统计学分析：数据以均数±标准差（±s）表示，采用SAS 8.0统计学软件进行t检验，P＜0.05为差异有显著性。
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